Single nucleotide polymorphism of miR-149 and susceptibility of rheumatoid arthritis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the association between microRNA (miR)-149 polymorphism and susceptibility to rheumatoid arthritis (RA ), as well as the clinical characteristics in patients with RA .@*METHODS@#A total of 200 RA patients and 120 healthy controls were recruited from Department of Rheumatology and Immunology of Nanjing First Hospital. After obtaining the informed consent, we collected 2 mL of anti-coagulated venous blood samples from all studied subjects to isolate the whole blood genomic DNA, and the clinical data were collected as well. Single nucleotide polymorphisms of miR-149 rs22928323 were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Correlation between single nucleotide polymorphisms and clinical features were compared.@*RESULTS@#The frequencies of TT, TC and CC for rs22928323 of miR-149 were 25.3%, 51.1% and 23.6% or 18.3%, 20.0% and 61.7% in the patients or the healthy controls, respectively. The onset risk of allele C in RA patients was increased compared with allele T [OR=1.38, 95% CI (1.01-1.75), P=0.023]. There were no significant difference in rheumatoid factor, blood urine nitrogen, antikeratin antibody, and other clinical characteristics among the 3 genotypes in RA patients (P>0.05).@*CONCLUSION@#SNP rs22928323 in miR-149 is correlated with RA in the east of Chinese Han population, whereas there is no correlation between miR-149 polymorphism and clinical characteristics in patients with RA.

Observation of mast cells and detection of related cytokines in murine bronchial asthma model pre and post desensitization therapy / 中国免疫学杂志

Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .

Biological characteristics of eutopic endometrial stromal cells in ovarian endometriosis / 中国综合临床

ObjectiveTo investigate eutopic endometrial stromal cells' ability of proliferation,apoptosis and invasion in ( ovarian endometriosis,OEMs).Methods Culture and identify eutopic endometrial stromal cells in 32 cases of patients with OEMs and 32 cases of patients with benign teratoma.Apply methyl thiazolyl tetrazolium methods,flow cytometric and transwell methods to detect these cells' capacity of proliferation,apoptosis and invasion.ResultsEutopic endometrial stromal cells were cultured successfully and their purity was more than 90％.The zero hour's absorbance of OEMs group was similar with control group (0.127 ±0.013vs 0.129 ±0.008,P ＞ 0.05).The growth of 24 and 48 hour's absorbance of OEMs group was significantly higher than control group (24 h -0 h:0.148 ±0.020 vs 0.048 ±0.008,t =26.066;48 h -24 h:0.397 ±0.029vs 0.119 ±0.022,t =42.544,P ＜0.01 ).The apoptosis rate of eutopic endometrial stromal cells in OEMs group was (26.430 ± 3.789 )％ and ( 35.571 ± 4.485 ) ％ in control group,which reached statistical difference ( t =- 8.808,P ＜0.01 ).After the eutopic endometrial stromal cells got through the small room,the absorbance in OEMs group(0.950 ± 0.014) was significantly higher than control group (0.653 ± 0.028 ) ( t =52.947,P ＜0.01 ).ConclusionThe proliferation and invasion capacity of eutopic endometrial stromal cell in OEMs group was more powerful than control group,however the apoptosis ability of this cell in OEMs group was weaker than control group.The change of biological characteristics of eutopic endometrial stromal cells of OEMs might be involved in the occurrance and development of OEMs.